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Label-free extraction of extracellular vesicles using centrifugal microfluidics

机译:使用离心微流控技术无标记提取细胞外囊泡

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摘要

Extracellular vesicles (EVs) play an important role as active messengers in intercellular communication and distant microenvironment modeling. Increasingly, these EVs are recognized as important biomarkers for clinical diagnostics. However, current isolation methods of EVs are time-consuming and ineffective due to the high diffusive characteristics of nanoparticles coupled with fluid flow instability. Here, we develop a microfluidic CEntrifugal Nanoparticles Separation and Extraction (µCENSE) platform for the rapid and label-free isolation of microvesicles. By utilizing centrifugal microhydrodynamics, we subject the nanosuspensions between 100 nm and 1000 nm to a unique fluid flow resulting in a zonal separation into different outlets for easy post-processing. Our centrifugal platform utilizes a gentle and efficient size-based separation without the requirements of syringe pump and other accessories. Based on our results, we report a high separation efficiency of 90% and an extraction purity of 85% within a single platform. Importantly, we demonstrate high EV extraction using a table top centrifuge within a short duration of eight minutes. The simple processes and the small volume requirement further enhance the utility of the platform. With this platform, it serves as a potential for liquid biopsy extraction and point-of-care diagnostics.
机译:细胞外囊泡(EV)在细胞间通讯和远距离微环境建模中作为主动信使发挥着重要作用。这些电动汽车越来越被认为是用于临床诊断的重要生物标志物。然而,由于纳米颗粒的高扩散特性以及流体流动的不稳定性,目前的电动汽车隔离方法既费时又无效。在这里,我们开发了一种微流体Centrifugal纳米颗粒分离和提取(µCENSE)平台,用于快速和无标签的微囊泡分离。通过利用离心微流体动力学,我们使100 nm至1000 nm之间的纳米悬浮液经受独特的流体流动,从而将区域性分离成不同的出口,以便于后处理。我们的离心平台利用基于大小的轻柔高效分离,而无需注射泵和其他附件。根据我们的结果,我们报告在一个平台上分离效率高达90%,提取纯度为85%。重要的是,我们演示了使用台式离心机在八分钟的短时间内实现高EV提取。简单的过程和小的体积要求进一步增强了平台的实用性。有了这个平台,它就有可能用于液体活检提取和即时诊断。

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